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Development of a gas chromatography/mass spectrometry based metabolomics protocol by means of statistical experimental design

Author:
  • Anders Danielsson
  • Thomas Moritz
  • Hindrik Mulder
  • Peter Spégel
Publishing year: 2012
Language: English
Pages: 50-63
Publication/Series: Metabolomics
Volume: 8
Issue: 1
Document type: Journal article
Publisher: Springer

Abstract english

Metabolomics is a growing research field where new protocols are rapidly developed and new applications discovered. Common applications include biomarker discovery and elucidation of drug metabolism. However, the development of such protocols rarely includes a systematic optimization followed by validation with real samples. Here a GC/MS-based protocol using methoximation followed by silylation with N-tert-butyldi-methylsilyl-N-methyltrifluoroacetamide (MTBSTFA) for analysis of blood plasma metabolites is thoroughly developed and optimized from derivatization to detection with statistical design of experiments (DOE). Validation was performed with blood plasma samples and proved the methodology to be efficient, rapid and reliable with a total of 51 analyses performed in 24 h, with linear responses, low detection limits and good precision. The obtained chromatograms were much cleaner, due to the absence of glucose overloading, and the data was found to drift less with MTBSTFA derivatisation than with MTBSTFA derivatisation.

Keywords

  • Endocrinology and Diabetes
  • Metabolomics
  • Gas chromatography
  • Mass spectrometry
  • MTBSTFA
  • Blood
  • plasma
  • DOE

Other

Published
  • Molecular Metabolism
  • ISSN: 1573-3882
Peter Spegel
E-mail: peter.spegel [at] chem.lu.se

Researcher

Centre for Analysis and Synthesis

1

Lund University Diabetes Centre, CRC, SUS Malmö, Entrance 72, House 91:12. SE-205 02 Malmö. Telephone: +46 40 39 10 00